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Steroids are often used in topical ointments due to their anti-inflammatory and immunosuppressive properties. However, their use may also result in various side effects. As such, it is important to accurately measure the concentration of steroids in topical ointments. Presented here is a method for analysis of steroids in commercially available topical ointments using the highly flexible Hitachi Chromaster HPLC system with UV-vis detection.